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Interleukin 1b (IL-1b) is a member of the cytokine family that serves as major mediators for early pro-inflammatory responses. During artificial breeding of Pacific cod (Gadus macrocephalus), outbreaks of NNV in the early development stages could lead to high mortality. Recent research reported that IL-1b participated in the host-virus interaction against NNV infection, however, no IL-1b gene has been identified from Pacific cod so far. In this study, we described the identification and characterization of a IL-1b from Pacific cod transcriptome at both transcript and protein level. Sequence alignment and phylogenetic analysis were conducted to determine and confirm their evolutionary relationship. Expression analysis revealed that IL-1b was expressed in all tested tissues and the western blot analysis confirmed the same expression profiles using purified recombinant IL-1b protein. In addition, significant up-regulation was detected in IL-1b transcripts during early development stages at 30dph and 50dph. After challenged with poly I:C, IL-1b was significantly induced after 24h. Taken together, these results strongly suggested that IL-1b might play an essential role in innate defense mechanism of Pacific cod.The IL-1b gene of Pacific cod (Gadus microcephalus) was identified and characterized.Phylogenetic analysis indicated that Pacific cod IL-1b exhibited the closest relationship to Atlantic cod IL-1b.IL-1b was ubiquitously expressed in spleen, kidney, ovary, heart, liver, gill, muscle and intestine.Expression analysis showed that IL-1b transcript significantly increased after 30 and 50 day-post-hatch.Significant induction was detected after poly I:C challenge.